Disodium [6-[(2-anilino-1-naphthyl)azo]-2,4-dinitrophenolato(2-)][3-hydroxy-4-[(2-hydroxy-1-naphthyl)azo]naphthalene-1-sulphonato(3-)]chromate(2-)

Endpoint summary

Description of key information

Non skin irritating

Non eye irritating

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Housing: in sigle cages.
- Diet: standard laboratory diet, ad libitum. The batch of diet used was analysed for chemical and microbiological contaminants.
- Water: ad libitum. The water bottles were withdrawn two hours prior and four hours after dosing.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 2 °C
- Humidity: 30 - 70 %
- Air changes: approximately 15 changes/hour.
- Photoperiod: 12 hrs dark / 12 hrs light.

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

- Amout: 1.0 ml
- Vehicle: the substance was used diluted in 50 % water.

Duration of treatment / exposure:

4 hour

Observation period:

72 hours

Number of animals:

Three male rabbits.

Details on study design:

TEST SITE
- Area of exposure: 24 hours prior to the test, the skin on the back of the animals was clipped by means of an electric clipper and the skin was investigated for pre-existing injuries. Test item was introduced under a square patch of surgical gauze measuring 6.5 cm2.
- Type of wrap if used: the animals were immobilized with patches secured in place by a plastic patch and the entire trunk was wrapped with a plasic sIeeve for the 4 hour exposure.

SCORING SYSTEM:
The irritation was measured by patch test technique on intact skin.
After 4 hours, the patches were removed and the resulting reactions were evaluated on the basis of the code of Federal Regulations, Title 16, Section 1500.41.

Irritation parameter:

erythema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2.3

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2.3

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The irritation score was 0.0
The substance can be considered as non irritating, according to the ETAD-Subcommittee for Toxicology and also according to the EC-Regulation.

Animal	Reaction	30 min	24 hrs	48 hrs	72 hrs
6 male	Erythema	0	0	0	0
	Oedema	0	0	0	0
7 male	Erythema	0	0	0	0
	Oedema	0	0	0	0
8 male	Erythema	0	0	0	0
	Oedema	0	0	0	0

Interpretation of results:

other: not classified, according to the CLP Regulation (EC) No 1272/2008

Conclusions:

Non irritating

Executive summary:

Three male rabbits were used to test the skin irritation potential of the substance. The skin of rabbits was treated with 1.0 ml of test item, which was used diluted in 50 % water. The irritation was measured by patch test technique on the intact skin. After 4 hours, the patches were removed and the resulting reactions were evaluated on the basis of the code of Federal Regulations, Title 16, Section 1500.41.

None of the animals showed any observable response to treatment throughout the 72 hours observation period.

The irritation score was 0.0 and the substance has been judged to be non irritating, according to the ETAD-Subcommittee for Toxicology and also according to the EC-Regulation.

Conclusion

The scoring system criteria used to record the hardness of the skin reactions followed in the current test are the same as those reported into the OECD guideline, therefore it is possible to re-evaluate the results in accordance with the CLP Regulation (EC) No 1272/2008.

None of the animals showed any observable response to treatment throughout the 72 hours observation period, thus it can be concluded that the test item does not meet the criteria to be classified as irritating, according to the CLP Regulation (EC) No 1272/2008.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

not specified

Species:

rat

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Housing: in sigle cages.
- Diet: standard laboratory diet, ad libitum. The batch of diet used was analysed for chemical and microbiological contaminants.
- Water: ad libitum. The water bottles were withdrawn two hours prior and four hours after dosing.
- Acclimation period: 5 days.
- Health check: prior to the test, the eyes of each rabbit were exami ned for pre-existing casual injury.

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 2 °C
- Humidity: 30 - 70 %
- Air changes: approximately 15 changes/hour.
- Photoperiod: 12 hrs dark / 12 hrs light.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 mg of test material was placed into the conjunctival sac.

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

Three male rabbits

Details on study design:

SCORING SYSTEM
The eyes were inspected after 30 minutes, 24, 48 and 72 hours.
Reading scores for ocular lesions were made according to the recommendations of the USA Food and Drug officials, published in 1959 (Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics).

Irritation parameter:

cornea opacity score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal:

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

chemosis score

Basis:

animal:

Time point:

24/48/72 h

Score:

< 2

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

The irritation score resulted to be 0.83.
The substance can be considered as non irritating, according to the ETAD-Subcommittee for Toxicology and also according to the EC-Regulation.

Summary of ocular findings

Animal	Reaction	30 min	24 hrs	48 hrs	72 hrs	Mean 24/48/72 hrs
1 male	Corneal opacity	0	0	0	0	0.00
	Iris	0	0	0	0	0.00
	Conjunctival redness	0	1	1	0	0.67
	Conjunctival chemosis	0	0	0	0	0.00
2 male	Corneal opacity	0	0	0	0	0.00
	Iris	0	0	0	0	0.00
	Conjunctival redness	0	1	1	0	0.67
	Conjunctival chemosis	0	0	0	0	0.00
3 male	Corneal opacity	0	0	0	0	0.00
	Iris	0	0	0	0	0.00
	Conjunctival redness	0	1	0	0	0.33
	Conjunctival chemosis	0	0	0	0	0.00

Reactions scored

Animal	Reaction		30 min	24 hrs	48 hrs	72 hrs
1 male	Cornea	(A) dendity	0	0	0	0
		(B) area	0	0	0	0
		A×B×5	0	0	0	0
	Iris	(C)	0	0	0	0
		C×5	0	0	0	0
	Conjunctiva	(D) secretion	0	0	0	0
		(E) swelling	0	0	0	0
		(F) redness	0	1	1	0
		(D+E+F)×2	0	2	2	0
2 male	Cornea	(A) dendity	0	0	0	0
		(B) area	0	0	0	0
		A×B×5	0	0	0	0
	Iris	(C)	0	0	0	0
		Cx5	0	0	0	0
	Conjunctiva	(D) secretion	0	0	0	0
		(E) swelling	0	0	0	0
		(F) redness	0	1	1	0
		(D+E+F)x2	0	2	2	0
3 male	Cornea	(A) dendity	0	0	0	0
		(B) area	0	0	0	0
		AxBx5	0	0	0	0
	Iris	(C)	0	0	0	0
		C×5	0	0	0	0
	Conjunctiva	(D) secretion	0	0	0	0
		(E) swelling	0	0	0	0
		(F) redness	0	1	0	0
		(D+E+F)×2	0	2	0	0
Score 10 : 12 = 0.83			0	6	4	2

Interpretation of results:

other: not classified, according to the CLP Regulation (EC) No 1272/2008

Conclusions:

Non irritating

Executive summary:

Three male rabbits were used to test the eye irritation potential of the substance. 100 mg of undiluted test material were placed into the conjunctival sac. The eyes were inspected after 30 minutes, 24, 48 and 72 hours. Reading scores for ocular lesions were made according to the recommendations of the USA Food and Drug officials, published in 1959 (Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics).

The irritation score resulted to be 0.83. The substance can be considered as non irritating, according to the ETAD-Subcommittee for Toxicology and also according to the EC-Regulation.

Conclusion

Although the criteria for the evaluation of the eye irritation potential followed in the test are different from those reported into the OECD guideline, the scoring system is the same. Raw tables are included into the study report, thus it is possible to re-evaluate the results according to the CLP Regulation (EC) No 1272/2008.

Mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for iritis, lower than 2 for both conjunctival redness and chemosis, in all tested animals, thus the substance does not meet the criteria to be classified as eye irritating, according to the CLP Regulation (EC) No 1272/2008.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

SKIN IRRITATION

The skin irritation potential of Acid Black 222 was assayed in two experiments, which were performed on test item with a different purity degree and dilution.

In both cases, three male rabbits were used and the irritation was measured by patch test technique on the intact skin. After 4 hours, the patches were removed and the resulting reactions were evaluated on the basis of the code of Federal Regulations, Title 16, Section 1500.41.

In both cases, none of the animals showed any observable response to treatment throughout the 72 hours observation period.

EYE IRRITATION

The eye irritation potential of Acid Black 222 was assayed in two experiments, which were performed on test item with a different purity degree and dilution.

Three male rabbits were used in both cases. The eyes were inspected after 30 minutes, 24, 48 and 72 hours and the reading scores for ocular lesions were made according to the recommendations of the USA Food and Drug officials, published in 1959 (Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics).

In both cases, no reactions were scorded for corneal opacity, iritis and conjunctival chemosis. In case of test item with an higher purity degree, conjunctival redness was recorded in all tested animals; anyway, the mean for gradings was lower than 2 and all the effects were fully reversible within 72 hours.

Justification for classification or non-classification

According to the CLP Regulation (EC) No 1272/2008, 3.2 Skin corrosion/irritation section, skin irritation means the production of reversible damage to the skin following the application of a test substance for up to 4 hours.

The mean values from gradings at 24, 48 and 72 hours after patch removal were lower than 2.3 in all animals for both erythema/eschar and oedema reactions.

According to the CLP Regulation (EC) No 1272/2008, serious eye damage means the production of tissue damage in the eye, or serious physical decay of vision, following application of a test substance, which is not fully reversible within 21 days of application.

The mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for iritis, lower than 2 for both conjunctival redness and oedema, in all the tested animals. In addition, all the recorded effects were fully reversible within 72 hours.

In conclusion, the substance does not meet the criteria to be classified for eye/skin irritation, according to the CLP Regulation (EC) No 1272/2008.